Exclusion of HLA-C Genotype with Zero Mismatched PCR-SBT Results by Next Generation Sequencing.
10.19746/j.cnki.issn.1009-2137.2022.04.038
- Author:
Yan-Ping ZHONG
1
;
Hao CHEN
1
;
Dan ZHOU
1
;
Hong-Yan ZOU
2
Author Information
1. Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center, Shenzhen 518020, Guangdong Province, China.
2. Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center, Shenzhen 518020, Guangdong Province, China E-mail: 958567432@qq.com.
- Publication Type:Journal Article
- Keywords:
Next-Generation Sequencing;
PCR-sequence-based typing;
human leukocyte antigen;
novel allele;
rare allele
- MeSH:
Alleles;
Genotype;
HLA-C Antigens/genetics*;
High-Throughput Nucleotide Sequencing;
Histocompatibility Testing/methods*;
Humans;
Polymerase Chain Reaction/methods*;
Sequence Analysis, DNA
- From:
Journal of Experimental Hematology
2022;30(4):1213-1218
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:Three cases of rare alleles of HLA-C with zero mismatched PCR-SBT results were analyzed by full-length sequencing to determine the true genotypes.
METHODS:Three rare HLA-C alleles with zero mismatched PCR-SBT results were screened from clinical transplant matching samples, and the full-length sequence was detected by next-generation sequencing technology.
RESULTS:The results of PCR-SBT typing of 3 samples were: HLA-C*03:04, 12:167; HLA-C*07:291, 15:02; HLA-C*01:43, 08:16. Other alleles were not in the CWD table of common and confirmed HLA alleles in China (version 2.3) except common allele HLA-C*03:04, HLA-C*15:02. NGS full-length sequencing revealed that the HLA-C genotypes of the three samples were a combination of common alleles and novel alleles, and the three novel alleles had a base mutation in exons 6, 2, and 4, respectively. The novel allele sequences have been submitted to the Genbank database (MK629722, MK335474, MK641803), which were officially named HLA-C*03:04:74, HLA-C*15:192, HLA-C*08:01:25 by the WHO HLA Nomenclature Committee. The HLA high-resolution typing results of 3 samples were: HLA-C*03:04:74, HLA-C*12:03; HLA-C*07:02, HLA-C*15:192; HLA-C*01:02, HLA-C*08:01:25.
CONCLUSION:HLA typing results containing rare alleles should be treated cautiously, and the full-length sequence should be verified by NGS or cloning. The laboratory finally confirmed that the 3 cases of PCR-SBT zero mismatch HLA-C genotypes are the combination of common alleles and novel alleles by NGS sequencing, which provides an accurate basis for clinical transplantation matching and enriches the human HLA genetic database.
